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OfflineLectric
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Agar Agar Powder w/ SUGAR?
    #27305559 - 05/12/21 10:24 AM (3 years, 7 days ago)

finally found Agar Agar powder but noticed it also has sugar in it.

will that still work for agar plates?

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InvisibleMilk-away
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Re: Agar Agar Powder w/ SUGAR? [Re: Lectric]
    #27306659 - 05/13/21 06:26 AM (3 years, 6 days ago)

technically.. yes, but it is not recommended.. white processed sugar (sucrose) is a very simple compound, so every organism can easily process it..
Your mycelium may turn wispy and slow, due to this, it's better to give it more complex nutrients, like yeast (micronutrients) and molasses(brown sugar), instead of white sugar.

All in all, you can use this agar in case it doesn't contain other ingredients (like vitamin C, or some fats), just make sure to take into account the extra sugar you are adding.. e.g: if your powder is 10%sugar, and recipe requires 10g of agar, you'd have to add 11.11g agar powder and take into account that 1g of sugar has already been added, so if recipe requires 10g honey, you'd only add 9.. etc

Good luck :thumbsup2:


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OfflineLectric
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Re: Agar Agar Powder w/ SUGAR? [Re: Milk-away] * 1
    #27307128 - 05/13/21 02:31 PM (3 years, 6 days ago)

awesome, thanks man!

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Invisibleninja cat 09
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Re: Agar Agar Powder w/ SUGAR? [Re: Lectric]
    #27307254 - 05/13/21 04:27 PM (3 years, 6 days ago)

Are there any other disadvantages to using sugar? I read that it can be used as a substitute for dextrose, but in Growing Gourmet and Medicinal Mushrooms, Stamets says something along the lines that simple sugars can make the mycelium "lazy" because it doesn't have to work for its food. Is this the case if I use it as a substitute for dextrose in PDA or PDYA?


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OfflineSolipsis
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Re: Agar Agar Powder w/ SUGAR? [Re: ninja cat 09] * 2
    #27315528 - 05/19/21 08:20 PM (3 years, 2 hours ago)

I think the way a lot of things are regulated in organisms can go by principle of 'use it or lose it', if it is not needed to toggle on genes that code for the production of enzymes needed to break down complex nutrients, preferably its turned off cause it costs resources.
Not entirely sure but i do also think that when something like that goes on for too long, somehow it can get more difficult, shocking or overwhelming if the myc is later transferred to a substrate that takes a lot more work to digest than what it "trained for", yea not sure how gene regulation "profiles" are "programmed" so to speak lol. 

Anyway it makes a lot more sense if a culture gets perfectly representative training for the work it will be doing later, right? it can adapt its metabolism nicely finetuned without having to do ridiculous reorganization at some point, haha.
It can be hard to avoid sometimes when we switch between types of media... LC put on agar can behave weirdly at first lol, wouldn't you say?

Actually recently I have been wondering about going even further than using grain soak/simmer water to make agar and optimize strains at an early stage (i don't mean making an isolate purely on agar, i am not really for that unless you screen a load of em).. beyond that, i thought having an actual puree of the grains you will later use as spawn (which is for cubes basically also your real substrate), that might be an ideal way to represent it. 
I assume there can be drawbacks like it may be a bit more work and quite possibly the growth is slower although its a matter of dilution, i def meant diluting that puree/suspension with water. 

As for your question there, ninja cat 09... I started out by explaining a bit about what that "laziness" mentioned may work like, and that would be when you use relatively too much of these simple sugars as your main carbs. But dextrose is actually an even simpler sugar than "table sugar" is.. Dextrose is a single sugar building block (saccharide) while table sugar or saccharose is composed of two connected sugar building blocks. 
PDA is not bad for having a simple sugar in it, not at all.. it also has a bunch of potato water in it which should have carbs that are a bit more complex, although of the most complex stuff: starch which is super long chains of sugar building blocks (i.e. polysaccharide), it has only quite limited solubility in water and its what makes it so milky clouded... i guess its starches on the edge of dissolving or something like that haha.

I always imagine it like shifting gears. It's good to include a mix of carbs of different lengths (so simple ones and complexer longer ones), I think because it should encourage a metabolism that at the very start of germination and growth can make swift use of the simple carbs present as startup resources, but there is a limit to what is present to there will be developed some sort of strategy/adaptation to use the energy gained by starting up cheaply, to use that available energy for investing in breaking down the complexer starches so that it can sustain itself.
That way you can try to find a balance between helping a hand with the addition of the free simple sugars, but while keeping the nutritional profile similar enough to what (grains, but can also be wood or poop and straw) you are actually gonna grow on.

So I would say that is the full story and seems to me it means you should not substitute carbs for one another in a way that might make your myc grow without having to invest energy of its own, but that prepares it poorly for real life as a grown up fungus on a substrate.
The difference between dextrose and saccharose is i think not such a big deal, its more like think about grains that are chockful of starch, very long chains... makes dextrose and saccharose difference insignificant but i think if you have the choice you would pick dextrose as your ultimate cheap startup carb. Doesn't mean that if you have saccharose at home but not dextrose and you wanna make agar now and not have that hassle, that it shouldnt be perfectly fine to use saccharose.

I heard some vague rumor about supposedly fungi having issues breaking down maltodextrin which is a pretty interesting intermediate length carb - but i think something misunderstood something there quite out of context. Sugars i think is one of the ultimate bounties in nature lol, hard currency.. probably also why its pretty addictive.. i would be a bit surprised if species are unable to break down polysaccharides (of certain length, or whatever, slightly less so sure).. unless somehow that fungus leads a life where it manages to never find maltodextrin or starch level complex carbs... :P

Another thing that fascinates me in principle is modified starch, which can often mean it is chemically altered to be water soluble which regular starch is not... i wonder if that would make the average fungus - say in LC - flip out totally cause of the impossibly high levels in solution, or rather if myc wouldn't really give a shit cause to it the difference between starch in suspension and in solution is in practice too limited heh not sure. Still you can probably have considerably more in solution than manages to stay in suspension without crashing out. well could be a fun experiment.. and seems like a non-opaque PDA version could be pretty sexy.
Anyway, how much it would behave as if it was itself agar, and if the modification makes it totally impossible for many mycs to eat the modified starch... those are new questions that arise. Maybe its not too different from say kappa-carrageenan.

well have fun with your carbs, its a cool n useful world

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Invisibleninja cat 09
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Re: Agar Agar Powder w/ SUGAR? [Re: Solipsis]
    #27316143 - 05/20/21 10:21 AM (2 years, 11 months ago)

Thanks for going into such depth! If I understood you correctly, then mycelium might like potato flake agar more than filtered potato tea, right?

I tried adding some wood to my agar, like Stamets suggests in his book, but I had the issue that most of it sank to the bottom of the bottle/plates, so only some plates got it and even then I only think it'll be useful for thinner plates. We'll see how this works out.


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OfflineSolipsis
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Re: Agar Agar Powder w/ SUGAR? [Re: ninja cat 09]
    #27320860 - 05/24/21 06:25 AM (2 years, 11 months ago)

It depends what you wanna do at what stage.. :smile:  There can be a paradox: if the myc likes something it often grows slower (i mean in exploration rather than growing in density) cause it stops looking so hard.

I guess it's not all that common practice what I suggest but yea if you are selecting strains from MS germination that seems pretty good, but if you only want to make a transfer of an established culture later for other reasons or you just wanna get a plate colonized asap to inoculate spawn with the agar then it seems better to use more limited nutes and of more limited complexity cause you just wanna get some ground covered at that point.

Yea i have had sawdust sitting on the bottom of my agar before, not sure i think the rest of the agar was a lower concentration LME or something? anyway from what i vaguely remember it worked surprisingly well but not sure how much the sawdust had to do with that.

I didn't realize it would sink so much either when i tried, now knowing this i think i would soak the sawdust much longer so that it sinks before PCing so that the amount can be chosen so that its level - if that ends up making it difficult (from a quick test) to pour equal amounts of sawdust with liquid, then maybe it is better to have an extra separate vessel with agar with no sawdust so that you can pour the sawdust first and then top it off?

I'm gonna discuss it some more on Discord, see if ppl agree that on paper it could or should be a superior way to get best substrate-optimized strains.

One question is if it's just way too early for that on like your first plate, also depending on how much spore material you apply and how much germinates. If there is a lot and you do a few transfers that also bring down the number of strains to a reasonable level, you could argue there are still enough strains to select from. But how big is the downside with having a part complex end substrate present right from the start? How limited/low do you need to make the simple sugars to force the myc to actually make use of that complex end substrate?
Could be challenging and it could also look discouraging if your start of growth can be slower than what we normally do.

IDK what species you were talking about btw with the wood in the agar?

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Invisibleninja cat 09
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Re: Agar Agar Powder w/ SUGAR? [Re: Solipsis]
    #27321221 - 05/24/21 01:44 PM (2 years, 11 months ago)

I think what would work best is to use smaller bottles of agar (400ml was too big) and add the final medium right before PCing, swirlingbefore pouring and between every one or two plates.

Stamets suggests using a normal agar recipe with about 3 grams/liter of final substrate added, but it would be interesting to see how the ratios affect the selection for a more substrate-optimized strain from spore.

In my case there's a few species, but I'm growing them from culture, since apparently it can also be benificial in that case to have final substrate in the petris. I'm growing a pair of oyster types (blue & pink), reishi, lions mane and shiitake. I'm using pine sawdust since it's very difficult to get hardwood sawdust here, there just isn't the market for it because most people here simply can't afford hardwoods.


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OfflineSolipsis
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Re: Agar Agar Powder w/ SUGAR? [Re: ninja cat 09]
    #27324533 - 05/27/21 01:25 AM (2 years, 11 months ago)

Quote:

ninja cat 09 said:
I think what would work best is to use smaller bottles of agar (400ml was too big) and add the final medium right before PCing, swirlingbefore pouring and between every one or two plates.

Stamets suggests using a normal agar recipe with about 3 grams/liter of final substrate added, but it would be interesting to see how the ratios affect the selection for a more substrate-optimized strain from spore.

In my case there's a few species, but I'm growing them from culture, since apparently it can also be benificial in that case to have final substrate in the petris. I'm growing a pair of oyster types (blue & pink), reishi, lions mane and shiitake. I'm using pine sawdust since it's very difficult to get hardwood sawdust here, there just isn't the market for it because most people here simply can't afford hardwoods.




Why do you think that is best? You think that sawdust might still be floating if it is only in the water during the PC cycle? Maybe, idk tho.. the heat and bubbling etc should speed up the hydration of the wood considerably but dont know how much. Also don't know if having the sawdust actually still floating is ideal cause it would ruin your nice smooth surface.
Having the sawdust right under the surface seems great, so that all the nice qualities of the gel can be used if you put spores on. Good hydration of the spores for example. I think its all good as long as the sawdust is relatively easily accessible, the myc should have no trouble finding it when exploring.
But anyway certainly worth testing what you suggest.

Its also easy to lose some moisture over time, seems best if it is lost from the agar surface without really affecting too much. If the sawdust or other sub material is too superficial it can make it difficult to see what you are doing regarding strain selection, sectoring etc. If its too deep, idk maybe its not even much of an issue.

Ah i see about the hardwood, there do exist fuel pellets with it (i got oak) but i guess it depends what is locally available and what people will buy. Well pine seems fine as long as it isn't quite fresh but aged enough. I heard you can also quickly treat sawdust instead of aging it, by boiling it a couple times with soda (refreshing the water used). A friend is working on almost the opposite of that though: he wants to optimize for what can grow on fresher pine, so then its like a bit of exposure therapy. :laugh:

I was thinking about working with a puree of grain spawn (for when the grain spawn is also basically the substrate), but from us talking about the sawdust, now i am actually wondering about having intact grains in the agar haha.

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Invisibleninja cat 09
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Re: Agar Agar Powder w/ SUGAR? [Re: Solipsis]
    #27325013 - 05/27/21 09:48 AM (2 years, 11 months ago)

I've been using small, granule like sawdust, so it hydrates very very easily with hot water, I don't think floating would be a problem, especially with longer 30 minutes PC cycles. The problem I had was clumping, but it might have been because I was pouring a bit too cold, hopefully future runs don't have that issue.

I've looked around for national suppliers for pellets and was only able to find one, and even then it was mostly pine wood being used.

I'd love to get something like that going, just using what's locally available as to reduce waste and costs. I don't know if they age the pine they use here or not, I don't think they do. I read a patent on Koreans using orange peels as a substrate up to 50% I think but they had the oils extracted for essential oil production, so I'm not sure if raw orange peels would work. I also saw people using coffee grains at about 70% sustrate, there's a lot of options if sawdust doesn't work, and I'm having fun experimenting and learning.

The grain idea might be a good one, but I've read that people use powdered grain and grain water with success. I have a bottle of grain water agar I prepped with about 200ml grain water, 200ml water, 8 grams agar, .8 grams yeast and 8 grams sugar. Apparently how much grain water to use really depends on how concentrated it is, I used a kilo of birdseed (it's a bit different from what you can get in the US, I think) and used about 3 liters of water, but came up with a really thick gooey "water", not sure if because the grains burst or because there was flax seed and other unusual grains in there.

I'll tell you how that goes when I pour the plates, I'm running through plates fast AF because my sterile technique is still shit :lol: Sugar and yeast might not have been necessary, but I decided to modify the PDYA recipe the least possible.


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OfflineDINKLB3RG
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Re: Agar Agar Powder w/ SUGAR? [Re: Milk-away]
    #27333889 - 06/03/21 01:35 PM (2 years, 11 months ago)

I probably shouldn't tell the forum about potato flake sucrose agar that Ive used in a pinch...


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InvisibleCreonAntigone
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Re: Agar Agar Powder w/ SUGAR? [Re: DINKLB3RG] * 1
    #27351404 - 06/16/21 10:22 PM (2 years, 10 months ago)

Is there something to the fact that malt extract has some special qualities that make it better at isolating fungi vs bacteria? I read that somewhere but never read the evidence for it. I'm not sure why malt extract would be better for fungi than other sugars, but my myc seems to like it.

Edited by CreonAntigone (06/16/21 10:22 PM)

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InvisibleBaba Yaga
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Re: Agar Agar Powder w/ SUGAR? [Re: Lectric] * 1
    #27351554 - 06/17/21 12:23 AM (2 years, 10 months ago)

Quote:

Lectric said:
finally found Agar Agar powder but noticed it also has sugar in it.

will that still work for agar plates?





So how much sugar is actually in your agar powder. I bought some from a health store in a pinch and it had 3% sugar in it which is only 0.3g when
using 10g. Nothing to worry about.

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